scholarly journals Prediction of N-linked Glycoform Profiles of Monoclonal Antibody with Extracellular Metabolites and Two-Step Intracellular Models

Processes ◽  
2019 ◽  
Vol 7 (4) ◽  
pp. 227 ◽  
Author(s):  
Sha Sha ◽  
Zhuangrong Huang ◽  
Cyrus Agarabi ◽  
Scott Lute ◽  
Kurt Brorson ◽  
...  

Monoclonal antibodies (mAbs) are commonly glycosylated and show varying levels of galactose attachment. A set of experiments in our work showed that the galactosylation level of mAbs was altered by the culture conditions of hybridoma cells. The uridine diphosphate galactose (UDP-Gal) is one of the substrates of galactosylation. Based on that, we proposed a two-step model to predict N-linked glycoform profiles by solely using extracellular metabolites from cell culture. At the first step, the flux level of UDP-Gal in each culture was estimated based on a computational flux balance analysis (FBA); its level was found to be linear with the galactosylation degree on mAbs. At the second step, the glycoform profiles especially for G0F (agalactosylated), G1F (monogalactosylated) and G2F (digalactosylated) were predicted by a kinetic model. The model outputs well matched with the experimental data. Our study demonstrated that the integrated mathematical approach combining FBA and kinetic model is a promising strategy to predict glycoform profiles for mAbs during cell culture processes.

2021 ◽  
Vol 1885 (3) ◽  
pp. 032004
Author(s):  
Qichao Pan ◽  
Zuwei Zhang ◽  
Yaqiong Zhang ◽  
Yaopeng Zhang ◽  
Bo Zhu

1986 ◽  
Vol 144 (3) ◽  
pp. 150-151 ◽  
Author(s):  
Francis G. Bowling ◽  
David K.B. Fraser ◽  
Alan E. Clague ◽  
Darryl J. Morris ◽  
Alan Hayes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nathan Jeger-Madiot ◽  
Lousineh Arakelian ◽  
Niclas Setterblad ◽  
Patrick Bruneval ◽  
Mauricio Hoyos ◽  
...  

AbstractIn recent years, 3D cell culture models such as spheroid or organoid technologies have known important developments. Many studies have shown that 3D cultures exhibit better biomimetic properties compared to 2D cultures. These properties are important for in-vitro modeling systems, as well as for in-vivo cell therapies and tissue engineering approaches. A reliable use of 3D cellular models still requires standardized protocols with well-controlled and reproducible parameters. To address this challenge, a robust and scaffold-free approach is proposed, which relies on multi-trap acoustic levitation. This technology is successfully applied to Mesenchymal Stem Cells (MSCs) maintained in acoustic levitation over a 24-h period. During the culture, MSCs spontaneously self-organized from cell sheets to cell spheroids with a characteristic time of about 10 h. Each acoustofluidic chip could contain up to 30 spheroids in acoustic levitation and four chips could be ran in parallel, leading to the production of 120 spheroids per experiment. Various biological characterizations showed that the cells inside the spheroids were viable, maintained the expression of their cell surface markers and had a higher differentiation capacity compared to standard 2D culture conditions. These results open the path to long-time cell culture in acoustic levitation of cell sheets or spheroids for any type of cells.


1989 ◽  
Vol 25 (9) ◽  
pp. 806-812 ◽  
Author(s):  
Tarek Bisat ◽  
Terry R. Brown ◽  
Claude J. Migeon ◽  
Gary D. Berkovitz

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